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Synergistic regulation of erythropoietin receptor (EPO‐R) expression by sense and antisense EPO‐R transcripts in the canine lung
Author(s) -
Zhang Quiyang,
Zhang Jianning,
Moe Orson W.,
Hsia Connie C.W.
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.601.3
Subject(s) - biology , complementary dna , downregulation and upregulation , sense (electronics) , microbiology and biotechnology , erythropoietin , hek 293 cells , receptor , gene , endocrinology , chemistry , genetics
We had found that EPO‐R protein is upregulated in growing canine lungs following pneumonectomy (PNX) ( Am J Physiol 287:L1107, 2004). We also found both sense and anti‐sense EPO‐R transcripts (sEPO‐R and asEPO‐R) to be upregulated in post‐PNX remaining lungs. We hypothesized that sEPO‐R and asEPO‐R interactions modulate EPO signaling in lung growth. We cloned a canine asEPO‐R cDNA, fully complementary to the sense strand of EPO‐R gene starting from 2.5 kb 3′‐ to the sense stop codon, and extending into the 5′UTR of sEPO‐R transcript. The asEPO‐R transcript harbors two putative open reading frames but it is not clear if they translated. The asEPO‐R and sEPO‐R transcripts co‐localize to bronchiolar epithelium and alveolar capillary endothelium. In HEK 293 cells transfection with sEPO‐R (+FLAG tag) cDNA alone increased EPO‐R protein expression, detected by anti‐EPO‐R and anti‐FLAG. At a constant sEPO‐R cDNA level, co‐transfection with escalating levels of asEPO‐R cDNA further increased recombinant EPO‐R protein expression. Thus, both sEPO‐R and asEPO‐R transcripts contribute to in vivo EPO‐R upregulation in the remaining lung post‐PNX. These data demonstrate novel synergism between complementary sense‐antisense transcripts in response to physiological stimulation in a robust model of induced lung growth. Supported by NHLBI RO1 HL40070, 62873, 45716, 54060, DK48482, DK‐20543 and Simmons Family Foundation.